s boulardii (ATCC)
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s boulardii
S Boulardii, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/s boulardii/product/ATCC
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
S Boulardii, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/s boulardii/product/ATCC
Average 93 stars, based on 1 article reviews
Price from $9.99 to $1999.99
s boulardii - by Bioz Stars,
2024-10
93/100 stars
Images
1) Product Images from "Functional Heterologous Protein Expression by Genetically Engineered Probiotic Yeast Saccharomyces boulardii"
Article Title: Functional Heterologous Protein Expression by Genetically Engineered Probiotic Yeast Saccharomyces boulardii
Journal: PLoS ONE
doi: 10.1371/journal.pone.0112660
Figure Legend Snippet: Tinoofpression by aromyces boulardii Auxotrophicine ii to create an optimal probiotic drug delivery system.
Techniques Used: Generated, Expressing, Marker
Figure Legend Snippet: Yeast were grown overnight in YPD and diluted to 10 7 cells per well in a 96 well plate. OD 600 readings over 24 hours incubation at 37°C or 30°C indicate relative growth of wild type S. boulardii (WT S.b. ), S. cerevisiae laboratory haploid ( S.c. lab haploid), S. cerevisiae wild type haploid ( S.c. WT haploid), and S. cerevisiae diploid ( S.c. diploid). Lines represent the mean of duplicate experiments, with error bars depicting plus the standard error of the mean (SEM). Shading highlights growth of yeast strains relative to growth of W T S. boulardii at 37°C.
Techniques Used: Incubation
Figure Legend Snippet: Wild type (WT) S. boulardii and S. cerevisiae diploid, haploid RAD1 , and haploid rad1 were exposed to various doses of UV irradiation. Percent survival (CFU as a percentage of total cells irradiated and plated) was plotted at each dose (mean of n = 2 per strain per UV dose, with error bars depicting plus the standard error of the mean) to identify the dose of UV irradiation corresponding to 50% survival of WT S. boulardii cells. Greater than 100% survival was likely reached at some low UV doses due to cellular replication after irradiation.
Techniques Used: Irradiation
Figure Legend Snippet: (A) Flow diagram depicting the number of irradiated wild type (WT) S. boulardii cells, screened 5-FOA resistant colonies, and final number of S. boulardii ura3 − mutants obtained. (B) Growth of WT S. boulardii , S. boulardii Mutants 1–3 (M1, M2, M3), and ura3 − S. cerevisiae laboratory haploid was assessed by serial dilution and spotting on YPD, uracil − , and 5-FOA plates. (C) Growth of S. boulardii Mutants 1–3 relative to WT S. boulardii and ura3 − S. cerevisiae laboratory haploid at 37°C in liquid media lacking uracil. Lines represent the mean of duplicate experiments for each strain, with error bars depicting plus and minus the standard error of the mean (SEM). (D) Number of CFU able to grow on plates lacking uracil per 10 9 plated cells. Each bar depicts the mean of duplicate experiments with error bars depicting plus the SEM.
Techniques Used: Irradiation, Serial Dilution
Figure Legend Snippet: (A) Schematic showing the domain structure of Ura3 protein in regions surrounding the amino acid changes in S. boulardii ura3 − mutants. Ura3 substrate binding sites are shown in gray with arrows above (amino acids 37, 59–61, 91–100, 217, 235) and the active site as a black line with asterisk above (amino acid 93). The altered amino acid sites in the S. boulardii mutants are shown as purple (S81F in M2) and yellow (A160S in M1 and M3) lines with the changes indicated below. Homologous regions including the altered amino acids and the 20 surrounding residues in Homo sapiens , Mus musculus , Danio rerio , Drosophila melanogaster , Saccharomyces cerevisiae , and WT S. boulardii are depicted to show conservation of these residues. (B) Ribbon depiction of the S. cerevisiae Ura3 homodimer bound to the proposed transition state analog 6-hydroxyuridine 5′-phosphate (PDB ID: 1DQX) . The S. boulardii mutant single amino acid changes are noted in yellow (A160S in M1 and M3) and purple (S81F in M2). (C) Enlarged view showing the wild type serine residue at position 81. (D) Enlarged view showing the amino acid change to phenylalanine at position 81 in S. boulardii Mutant 2. (E) Enlarged view showing the wild type alanine residue at position 160. (F) Enlarged view showing the amino acid change to serine at position 160 in S. boulardii Mutants 1 and 3.
Techniques Used: Binding Assay, Mutagenesis
Figure Legend Snippet: Yeast were grown overnight in YPD and diluted to 10 7 cells per well in a 96 well plate. OD 600 readings were taken over 24 hours incubation at 37°C. Graphs depict growth of yeast strains at pH 2, pH 4, pH 8, 0.3% OxGall, and YPD (approximately pH 6). Yeast strains include wild type (WT) S. boulardii (A); S. cerevisiae strains laboratory haploid (B), diploid (C), and wild type haploid (D); and S. boulardii M1 (E), M2 (F), and M3 (G). This analysis shows that S. boulardii mutants maintain resistance to pH 4 and pH 8 as well as to 0.3% OxGall whereas S. cerevisiae strains laboratory haploid and diploid are sensitive to these conditions.
Techniques Used: Incubation
Figure Legend Snippet: (A) Wild type (WT) S. boulardii ; S. cerevisiae strains laboratory haploid, diploid, and wild type haploid; and S. boulardii M1, M2, and M3 were grown overnight in YPD and diluted to 5×10 7 cells/mL in fresh YPD. OD 600 readings were taken over 24 hours incubation in a vinyl anaerobic chamber maintained at 37°C. (B) Number of colony forming units (CFU) per mL for each yeast strain after 12 and 24 hours incubation in the vinyl anaerobic chamber. This analysis shows that WT S. boulardii and particularly S. boulardii Mutants 1–3 show superior growth in anaerobic conditions relative to S. cerevisiae strains.
Techniques Used: Incubation
Figure Legend Snippet: (A) Bright field and fluorescent images of ura3 − S. cerevisiae laboratory haploid ( S.c. ) and S. boulardii Mutant 2 (M2) either untransformed (Control) or transformed (+GFP) with a URA3 plasmid containing GFP. The GFP fluorescence is detected in the FITC channel. Corresponding differential interference contrast (DIC) images are also shown. Scale bars show 10 µm. (B) Representative flow cytometry plots of forward-scattered light (FSC) versus GFP fluorescence for untransformed (Control) and transformed (+GFP) S. cerevisiae laboratory haploid ( S.c. lab haploid) and S. boulardii Mutant 2 (M2) showing the percent of GFP positive cells in each population (n = 2 per strain). Transformed yeast were maintained in media lacking uracil prior to analysis. (C) Retention of URA3 plasmid and GFP expression was tested by comparing the percent of GFP positive cells of untransformed yeast (Control) relative to transformed yeast cultured in either selective media lacking uracil (URA − Glu), YPD (non selective media) for 4 hours (YPD 4 hr), or YPD for 24 hours (YPD 24 hr). Yeast strains analyzed include untransformed and transformed ura3 − S. cerevisiae laboratory haploid ( S.c. ) and S. boulardii Mutants 1–3 (M1, M2, M3). Median fluorescent intensity (MFI) of GFP positive cells in each population is also depicted, indicating there is no visible decrease in average GFP expression per cell after incubation in YPD for 4 or 24 hours. Bars depict the mean of two samples per strain per incubation condition.
Techniques Used: Mutagenesis, Transformation Assay, Plasmid Preparation, Fluorescence, Flow Cytometry, Expressing, Cell Culture, Incubation
Figure Legend Snippet: (A) Schematic depicting oral gavage experiments. C57BL/6 mice were gavaged with 100 µL containing either water, 10 8 CFU wild type S. boulardii (WT S.b. ), 10 8 CFU S. boulardii Mutant 2 (M2), or 10 8 CFU ura3 − S. cerevisiae laboratory haploid ( S.c. ). Peyer's patches, sites of antigen sampling and immune response generation in the gastrointestinal tract (reviewed in ), were harvested 4 hours post gavage and plated to detect viable CFU. (B) Images of typical plates from oral gavage experiments showing recovery of viable yeast from Peyer's patches. Samples from mice gavaged with WT S. boulardii , S. boulardii Mutant 2 transformed with URA3 plasmid, or S. cerevisiae laboratory haploid transformed with URA3 plasmid were plated on media lacking uracil. Samples from naïve mice were also plated on YPD media to detect any contaminating yeast unable to grow without uracil. (C) CFU per mouse recovered from Peyer's patches of mice orally gavaged with water (Naïve), WT S. boulardii (WT S.b. ), S. boulardii Mutant 2 (M2), or S. cerevisiae laboratory haploid ( S.c. ) (n = 20 mice per group). Lines show the mean CFU per mouse for each group. Two data points for S. boulardii Mutant 2 (87 and 110 CFU per mouse) are not depicted in order to allow better visualization of other data points. The mean without the two high points is 2.5 (shown in solid black line). The mean including the two points is 12.1 (shown in dotted line). (D) Representative flow cytometry plots of forward-scattered light (FSC) versus GFP fluorescence showing the percent of GFP positive cells among untransformed S. boulardii Mutant 2 (M2 control) and S. boulardii Mutant 2 that was transformed with a URA3 plasmid encoding GFP (M2+GFP) and subsequently recovered from murine Peyer's patches (26 total transformed S. boulardii M2 CFU recovered from Peyer's patches were assessed by flow cytometry).
Techniques Used: Mutagenesis, Sampling, Transformation Assay, Plasmid Preparation, Flow Cytometry, Fluorescence
